STR基因座等位基因阶梯制备方法尝试

提高PCR-STR分型技术的准确性。采用荧光标记dNTP掺入法,通过DNA自动测序仪分析不同等位基因片段的长度并确定其命名后,建立并比较扩增产物混合-纯化-重扩增法、琼脂糖凝胶电泳-切割回收纯化DNA-重扩增法、非变性聚丙烯酰胺凝胶电泳-切割回收纯化DNA-重扩增法等3种Ladder制备方法。扩增产物混合-纯化-重扩增法相对较简便、快速、经济。Ladder对照使基因分型数据化,判型准确,重复性好,有利于标准化的实现。

Study on the preparation of locus specific allelic ladder

To improve the accuracy of STR-PCR typing method. The size of different alleles of D19S253 were analysed using GeneScan2.1 software on ABI Prisrn377 DNA Sequencer. 3 methods were used to produce allele ladder: (1) mixing PCR products containing different alleles-purifying- reamplifying; (2) making mixture-AE (agarose gel electrophoresis)-extracting and purifying DNA from gel-reamplifying; (3) making mixture-PAGE (native polyacrylamide gel electrophoresis)- extracting and purifying DNA from gel-reamplifying. The first method is simpler and more economic than others. When using allelic ladders, the results of STR-typing are more accurate. The method is easy to type.